RESUMEN
The clinical impact of therapeutic interventions in Parkinson's disease is often measured as a reduction in OFF-time when the beneficial effects of the standard-of-care L-DOPA formulations wanes off. We investigated the pharmacodynamic interactions of augmentation therapy to standard-of-care using a quantitative systems pharmacology (QSP) model of the basal ganglia motor circuit, essentially a computer model of neuronal firing in the different subregions with anatomically informed connectivity, cell-specific expression of 17 different G-protein coupled receptors and corresponding coupling to voltage-gated ion channel effector proteins based on experimentally observed intracellular signaling. The calculated beta/gamma (b/g) power spectrum of the local field potentials in the subthalamic nucleus was previously calibrated on the clinically relevant Unified Parkinson's Disease Rating Scale (UPDRS). When combining this QSP model with PK modeling of different formulations of L-DOPA, we calculated the b/g fluctuations over a 16 h awake period and used a weighted distance from a specific threshold to determine the cumulative liability of OFF-Time. Prediction of OFF-time with clinical observations of different L-DOPA formulations showed a significant correlation. Simulations show that augmentation with the adenosine A2A antagonist preladenant reduces OFF-time with 6 min for carbidopa/levodopa 950 mg 5-times daily to 37 min for 100 mg L-DOPA - 3 or 5 times daily. Exploring delays between preladenant and L-DOPA intake did not improve the outcome. Hypothetical A2A antagonists with an ideal PK and pharmacology profile can achieve OFF-Time reductions ranging from 9.5 min with DuoDopa to 55 min with low dose L-DOPA formulations. Combination of the QSP model with PK modeling can predict the anticipated OFF-Time reduction of novel A2A antagonists with standard of care. With the large number of GPCR in the model, this combination can support both the design of clinical trials with new therapeutic agents and the optimization of combination therapy in clinical practice.
Asunto(s)
Levodopa , Enfermedad de Parkinson , Humanos , Levodopa/farmacología , Enfermedad de Parkinson/tratamiento farmacológico , Antiparkinsonianos/farmacología , Antiparkinsonianos/uso terapéutico , Farmacología en RedRESUMEN
The Japan Bioanalysis Forum Symposium was held on 12-14 February 2019 (Yokohama, Japan), in celebration of its 10th anniversary, and over 370 participants from pharmaceutical industries, contractors, academia and regulatory authorities from home and abroad came together in Yokohama. The 3-day symposium particularly aimed to foster collaboration with the scientists surrounding bioanalysts, according to the theme 'Open to the Public.' The symposium also included a broad range of pioneering programs, such as lectures by speakers from DMPK/metabolomics fields, discussions of future bioanalysis and poster presentations by publicly offered presenters as well as the regular ones we had organized. This report summarizes the major topics as a conference report.
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Pruebas de Química Clínica , Biomarcadores/análisis , Interacciones Farmacológicas , JapónRESUMEN
Aim: Appropriateness of anti-drug antibody (ADA) assay is critical for immunogenicity assessment of biopharmaceuticals. Although cut point setting in ADA assay has a large impact on the results, a standard statistical approach for its setting has not been well established. Methodology: In this multi-laboratory study, to elucidate factors influencing the cut point setting, we compared the statistical approaches and calculated cut points for multiple datasets of ADA assays using the individual procedure employed at each laboratory. Conclusion: We showed that outlier exclusion, false-positive rate and investigating data distribution have the greatest impact on both screening and confirmatory cut points. Our results would be useful for industry researchers and regulators engaged in immunogenicity assessment of biopharmaceuticals.
Asunto(s)
Anticuerpos/análisis , Productos Biológicos/inmunología , Bases de Datos Farmacéuticas/estadística & datos numéricos , Inmunoensayo/estadística & datos numéricos , Algoritmos , Anticuerpos/inmunología , Humanos , Inmunoensayo/métodos , Modelos Estadísticos , Proyectos de InvestigaciónRESUMEN
The ninth Japan Bioanalysis Forum symposium took place at tower hall Funabori, Tokyo, Japan, between 6 and 8 February, 2018. Bioanalytical scientists from the pharmaceutical industry, CROs, academia and regulatory bodies had many meaningful and relevant discussions on current topics of interest in bioanalysis. The 3-day symposium featured updated perspectives and experiences on regulated bioanalysis of small and large molecules, biomarker measurement and assessment of immunogenicity, as well as new areas of bioanalytical validation such as quantitative polymerase chain reaction(qPCR) and flow cytometry. There were over 260 participants from six countries, with 23 oral and 11 poster presentations, including the outcomes of Japan Bioanalysis Forum discussion groups. This report summarizes the major discussion topics from the conference.
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Técnicas de Química Analítica , Métodos Analíticos de la Preparación de la Muestra , Anticuerpos/análisis , Anticuerpos/uso terapéutico , Cromatografía Liquida , Descubrimiento de Drogas , Humanos , Japón , Espectrometría de MasasRESUMEN
This study was undertaken to evaluate the performance of anti-drug antibody (ADA) assays constructed by each participating company using common samples including ADA, drug and human serum. The ADA assays constructed by each company showed good sensitivity and precision for evaluation of ADA. Cut points for screening and confirmatory assays and assay selectivity were determined by various calculation methods. In evaluations of blind ADA samples, nearly similar results were obtained by the study companies in determinations of whether samples were positive or negative except at the lowest sample concentration (5 ng/mL). In measurement of drug tolerance, for almost samples containing ADA and drugs, more positive results were obtained in assays using acid dissociation compared to those without acid dissociation. Overall, the performance of ADA assays constructed by the 10 companies participating in this study was acceptable in terms of sensitivity and reproducibility for detection and evaluation of immunogenicity in both patients and healthy subjects. On the other hand, based on results for samples containing ADA and drugs, validity of results for ADA assays conducted without acid dissociation was less meaningful and more difficult to evaluate. Thus, acid dissociation was confirmed to be useful for improving drug tolerance.
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Anticuerpos Monoclonales Humanizados/sangre , Inmunosupresores/sangre , HumanosRESUMEN
We previously reported that KW-2449, (E)-1-{4-[2-(1H-Indazol-3-yl)vinyl]benzoyl}piperazine, a novel multikinase inhibitor developed for the treatment of leukemia patients, was oxidized to an iminium ion intermediate by monoamine oxidase B (MAO-B) and then converted to its oxo-piperazine form (M1) by aldehyde oxidase (AO). However, it was found that the significant decrease in the pharmacologically active metabolite M1 following repeated administration of KW-2449 in primates might hamper the effectiveness of the drug. The mechanism underlying this phenomenon was investigated and it was found that the AO activity was inhibited in a time-dependent manner in vitro under the co-incubation of KW-2449 and MAO-B, while neither KW-2449 nor M1 strongly inhibited MAO-B or AO activity. These results clearly suggest that MAO-B catalysed iminium ion metabolite inhibited AO, prompting us to investigate whether or not the iminium ion metabolite covalently binds to endogenous proteins, as has been reported with other reactive metabolites as a cause for idiosyncratic toxicity. The association of the radioactivity derived from 14 C-KW-2449 with endogenous proteins both in vivo and in vitro was confirmed and it was verified that this covalent binding was inhibited by the addition of sodium cyanide, an iminium ion-trapping reagent, and pargyline, a MAO-B inhibitor. These findings strongly suggest that the iminium ion metabolite of KW-2449 is highly reactive in inhibiting AO irreversibly and binding to endogenous macromolecules covalently.
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Aldehído Oxidasa/antagonistas & inhibidores , Indazoles/metabolismo , Indazoles/farmacología , Piperazinas/metabolismo , Piperazinas/farmacología , Proteínas/metabolismo , Aldehído Oxidasa/metabolismo , Animales , Isótopos de Carbono , Humanos , Macaca fascicularis , Monoaminooxidasa/metabolismo , Inhibidores de la Monoaminooxidasa/farmacología , Oxidación-Reducción , Pargilina/farmacología , Unión Proteica , Ensayo de Unión Radioligante , Cianuro de Sodio/farmacologíaRESUMEN
Immunogenicity assessment is an important issue for ensuring the safety and efficacy of therapeutic protein products. Although the reliability of the anti-drug antibody (ADA) assay is one of the key points, there are some difficulties in assessing its validity because the analytes are polyclonal antibodies with variable and unknown characteristics. To elucidate the points to consider for the ADA assay, a Japanese research group was established that discusses the issues raised on the immunogenicity assessment. In this review, we first introduce the current situation regarding the development and immunogenicity assessment of therapeutic protein products in Japan. We then present our current view and recommendations on the ADA assay by considering its unique features.
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Bioensayo/métodos , Productos Biológicos/inmunología , Humanos , JapónRESUMEN
(E)-1-{4-[2-(1H-Indazol-3-yl)vinyl]benzoyl}piperazine (KW-2449) is a novel multikinase inhibitor. During our clinical study, we found that KW-2449 is mainly metabolized to its oxo-piperazine form (M1). An inhibition study suggested that monoamine oxidase-B (MAO-B) oxidizes KW-2449 to an iminium (intermediate) and aldehyde oxidase (AO) then metabolizes the intermediate to M1. The conversion of KW-2449 to the iminium (intermediate) by MAO-B was confirmed by the formation of its cyanide adduct. This cooperative metabolic pathway by MAO-B and AO was newly identified in the metabolism of piperazine. The clearance of KW-2449 by MAO-B and AO in human was estimated based on the kinetic analysis with in vitro-in vivo extrapolation. The systemic clearance in human was similar to the calculated value, indicating that the extrapolation approach was applicable to KW-2449 metabolism. Finally, we found that (E)-3-amino-1-{4-[2-(1H-Indazol-3-yl)vinyl]benzoyl}-pyrrolidine (Compound A) as a stable compound against MAO-B and AO. The total body clearance of Compound A was reduced to one tenth of KW-2449, demonstrating that preventing the metabolism of MAO and AO led to more preferable pharmacokinetic profiles. As piperazine is often introduced to drug candidates to improve lipophilicity of the compound to get more hydrophilic nature, the results of this study provide useful information for future drug development.
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Aldehído Oxidasa/metabolismo , Iminas/metabolismo , Indazoles/química , Indazoles/metabolismo , Monoaminooxidasa/metabolismo , Piperazinas/química , Piperazinas/metabolismo , Animales , Sistema Enzimático del Citocromo P-450/química , Sistema Enzimático del Citocromo P-450/metabolismo , Perros , Hepatocitos/química , Hepatocitos/metabolismo , Humanos , Iminas/química , Iones/química , Iones/metabolismo , Macaca fascicularis , Microsomas Hepáticos/química , Microsomas Hepáticos/metabolismo , Mitocondrias/química , Mitocondrias/metabolismo , Monoaminooxidasa/química , Oxidación-Reducción , Piperazina , RatasRESUMEN
Determination of estrogens in plasma is important in evaluation of effects of some anticancer drugs, such as aromatase inhibitors. However, as reported previously, high performance liquid chromatography-radio immunoassay (HPLC-RIA) and liquid chromatography-tandem mass spectrometry (LC-MS/MS) with chemical derivatization require complicated sample preparation. In this study, a highly sensitive and simple method for determination of estrone (E1), 17beta-estradiol (E2) and estrone 3-sulfate (E1S) in human plasma has been developed. Following diethylether extraction from plasma, analytes were purified by immunosorbents and then determined by LC-MS/MS using electrospray ionization (ESI). Immunosorbents were prepared by immobilization of specific antibodies raised against each analyte onto solid support. Use of selective immunosorbents in sample preparation removed interference in plasma samples that would cause ionization suppression, and markedly improved the sensitivity of LC-MS/MS for these analytes, without derivatization. Calibration curves of each analyte showed good linearity and reproducibility over the range of 0.05-50pg/injection for E1, 0.2-50pg/injection for E2 and 0.05-300pg/injection for E1S, respectively. The mean values of lower limits of quantification (LLOQ) in human plasma corrected by recovery of deuterated estrogens (internal standard, I.S.) were 0.1892pg/mL for E1, 0.7064pg/mL for E2 and 0.3333pg/mL for E1S, respectively. These LLOQ values were comparable to those previous reported using HPLC-RIA and LC-MS/MS. Using this method, the normal levels of three estrogens in healthy female plasma (n=5) were determined. The mean values of E1, E2 and E1S were 38.0pg/mL (range 24.8-53.0), 34.3pg/mL (22.6-46.6) and 786pg/mL (163-2080), respectively. The immunoaffinity LC-MS/MS described here allows sensitive and accurate quantification of E1, E2 and E1S without laborious sample preparation.
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Cromatografía Líquida de Alta Presión/métodos , Estradiol/sangre , Estrona/análogos & derivados , Estrona/sangre , Técnicas de Inmunoadsorción , Espectrometría de Masas en Tándem/métodos , Anticuerpos Inmovilizados/metabolismo , Estradiol/inmunología , Estradiol/aislamiento & purificación , Estradiol/metabolismo , Estrona/inmunología , Estrona/aislamiento & purificación , Estrona/metabolismo , Femenino , Humanos , Reproducibilidad de los Resultados , Sensibilidad y EspecificidadRESUMEN
Steroid sulfatase (STS) catalyses the hydrolysis of the sulfate esters of 3-hydroxy steroids, which are inactive transport or precursor forms of the active 3-hydroxy steroids. STS inhibitors are expected to block the local production and, consequently to reduce the active steroid levels; therefore, they are considered as potential new therapeutic agents for the treatment of estrogen- and androgen-dependent disorders such as breast and prostate cancers. KW-2581 is a novel steroidal STS inhibitor. In the present study, we found KW-2581 inhibited recombinant human STS (rhSTS) activity with an IC(50) of 2.9 nM when estrone sulfate was used as a substrate. The potency of KW-2581 was approximately 5-fold higher than that of a non-steroidal STS inhibitor, 667 COUMATE. KW-2581 was able to equally inhibit rhSTS activity when dehydroepiandrosterone sulfate was used as another substrate. KW-2581 inhibited rhSTS activity in a time- and concentration-dependent manner (k(inact), 0.439 min(-1); K(i, app), 15 nM), suggesting that it is an active site-directed irreversible inhibitor. Both decrease of KW-2581 concentration and increase of the des-sulfamoylated form's concentration were simultaneously observed during the reaction in a time-dependent manner with corresponding to the decrease of STS activity. Our findings for the first time demonstrated the production of des-sulfamoylated form of the compound as a consequence of STS inactivation.